Invitrogen

ARF6 Polyclonal Antibody

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated.

5 Published Figures

8 References

View all (14) ARF6 antibodies

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Protocols

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ARF6 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of ARF6 was performed using 70% confluent log phase PC-3 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with ARF6 Polyclonal Antibody (Product # PA1-093) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 40X magnification.

ARF6 Antibody in Immunohistochemistry (IHC) — Immunohistochemistry was performed on normal biopsies of deparaffinized human heart tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a Rabbit Polyclonal Antibody recognizing Arf6 (Product # PA1-093) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

ARF6 Antibody in Western Blot (WB) — Western Blot was performed using Anti-ARF6 Polyclonal Antibody (Product # PA1-093) and a 25 kDa band corresponding to ARF6 was observed across cell lines with uncharacterized band(*). Whole cell extracts (30 µg lysate) of LNCaP (Lane 1), MDA-MB-231 (Lane 2), HUVEC (Lane 3), MCF7 (Lane 4), MCF 10A (Lane 5), SK-BR-3 (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The Blot was probed with the primary antibody ( 1 mg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

ARF6 Antibody in Western Blot, Immunocytochemistry (WB, ICC/IF) — Figure 3 Secretion of sFlt1 from endothelial cells requires functional Arf6, Arf1, and Rab11 GTPases. A-F : EC-sFlt1 cells were nucleofected with plasmid constructs expressing dominant-negative forms of Arf1, Arf6, Rab5, Rab6, Rab 8, syntaxin 4, syntaxin 6, and syntaxin 16. Similarly, in a separate study EC-sFlt1 cells were nucleofected with scramble oligonucleotide or siRNAs against Arf1 , Arf6 , and Rab11a . Media containing nucleofection complexes were replaced with fresh media 8 hrs post-nucleofection. At 36 hrs post-nucleofection, secreted and intracellular sFlt1 were immunoprecipitated from spent culture media and cell lysates using the anti-v5 Ab. ( A , C ) Representative blot shows relative expression of sFlt1-v5 in cell lysates and secreted in culture media. B , D : Band densities from experiments as in A, C were quantified by densitometric analysis. Secretion of sFlt1-v5 was calculated as the ratio of desitometric band intensity values of secreted to intracellular Flt1; this ratio was set at 1 for mock-transfected control EC-sFlt1 cells. The data are the means +- SEM from three independent experiments. E : EC-sFlt1 cells were transiently transfected with indicated dominant-negative plasmid constructs. After 24 hrs cells were fixed, permeabilized, and labeled with anti-v5 Ab to detect cell-associated sFlt1-v5. Representative images obtained by epifluorescence microscopy show localization of sFlt1-v5. F : Quantification of intracellular retention of sFlt1-v5 upon inhi

ARF6 Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. MDA-MB-231 cells were transfected with ARF6 siRNA and decrease in signal intensity was observed in Western Blot application using Anti-ARF6 Polyclonal Antibody (Product # PA1-093). {KD}

Product Details

PA1-093

Applications	Tested Dilution	Publications
Western Blot (WB)	1:1,000	View 4 publications 4 publications
Immunohistochemistry (Paraffin) (IHC (P))	1:20-100	-
Immunocytochemistry (ICC/IF)	1:20-1:100	View 3 publications 3 publications
Miscellaneous PubMed (Misc)	-	View 1 publication 1 publication

Product Specifications
Species Reactivity	Human, Mouse, Non-human primate
Published species	Human, Mouse
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	C-terminal human recombinant protein fragment View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Protein A
Storage buffer	PBS with 1mg/mL BSA, 30% glycerol
Contains	0.05% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_2539860

Target Information

ARF6 (ADP-ribosylation factor 6) is a broadly expressed, differentially regulated GTPase that is present in cytoplasm and on both cell-surface and intracellular membranes and whose functions may include tissue-specific effects on vesicular trafficking during cellular differentiation.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

SARS-CoV-2 pseudovirus enters the host cells through spike protein-CD147 in an Arf6-dependent manner.

Emerging microbes & infections

Zhou YQ,Wang K,Wang XY,Cui HY,Zhao Y,Zhu P,Chen ZN

Published figure using ARF6 polyclonal antibody (Product # PA1-093) in Immunocytochemistry

Thu Dec 01 00:00:00 EST 2022

A mosquito salivary protein promotes flavivirus transmission by activation of autophagy.

Nature communications

Sun P,Nie K,Zhu Y,Liu Y,Wu P,Liu Z,Du S,Fan H,Chen CH,Zhang R,Wang P,Cheng G

Published figure using ARF6 polyclonal antibody (Product # PA1-093) in Immunocytochemistry

Tue Jan 14 00:00:00 EST 2020

Heterozygous loss of function of IQSEC2/Iqsec2 leads to increased activated Arf6 and severe neurocognitive seizure phenotype in females.

Life science alliance

Jackson MR,Loring KE,Homan CC,Thai MH,Määttänen L,Arvio M,Jarvela I,Shaw M,Gardner A,Gecz J,Shoubridge C

PA1-093 was used in Western Blotting to confirm the loss of Iqsec2 mRNA expression and lack of Iqsec2 protein within the brain of both founder and progeny mice.

Thu Aug 01 00:00:00 EDT 2019

Syndecan 1 is a critical mediator of macropinocytosis in pancreatic cancer.

Nature

Yao W,Rose JL,Wang W,Seth S,Jiang H,Taguchi A,Liu J,Yan L,Kapoor A,Hou P,Chen Z,Wang Q,Nezi L,Xu Z,Yao J,Hu B,Pettazzoni PF,Ho IL,Feng N,Ramamoorthy V,Jiang S,Deng P,Ma GJ,Den P,Tan Z,Zhang SX,Wang H,Wang YA,Deem AK,Fleming JB,Carugo A,Heffernan TP,Maitra A,Viale A,Ying H,Hanash S,DePinho RA,Draetta GF

PA1-093X was used in Western Blotting to investigate if a mechanistic link between KRAS* signaling and a targetable molecule driving nutrient salvage pathway in pancreatic ductal adenocarcinoma exists.